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• W2162007127 abstract "Francisella tularensis, the etiologic agent of tularemia in humans, is a potential biological threat due to its low infectious dose and multiple routes of entry. F. tularensis replicates within several cell types, eventually causing cell death by inducing apoptosis. In this study, a modified Himar1 transposon (HimarFT) was used to mutagenize F. tularensis LVS. Approximately 7,000 Km(r) clones were screened using J774A.1 macrophages for reduction in cytopathogenicity based on retention of the cell monolayer. A total of 441 candidates with significant host cell retention compared to the parent were identified following screening in a high-throughput format. Retesting at a defined multiplicity of infection followed by in vitro growth analyses resulted in identification of approximately 70 candidates representing 26 unique loci involved in macrophage replication and/or cytotoxicity. Mutants carrying insertions in seven hypothetical genes were screened in a mouse model of infection, and all strains tested appeared to be attenuated, which validated the initial in vitro results obtained with cultured macrophages. Complementation and reverse transcription-PCR experiments suggested that the expression of genes adjacent to the HimarFT insertion may be affected depending on the orientation of the constitutive groEL promoter region used to ensure transcription of the selective marker in the transposon. A hypothetical gene, FTL_0706, postulated to be important for lipopolysaccharide biosynthesis, was confirmed to be a gene involved in O-antigen expression in F. tularensis LVS and Schu S4. These and other studies demonstrate that therapeutic targets, vaccine candidates, or virulence-related genes may be discovered utilizing classical genetic approaches in Francisella." @default.
• W2162007127 created "2016-06-24" @default.
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• W2162007127 date "2007-11-01" @default.
• W2162007127 modified "2023-09-23" @default.
• W2162007127 title "Identification of <i>Francisella tularensis Himar1</i> -Based Transposon Mutants Defective for Replication in Macrophages" @default.
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• W2162007127 doi "https://doi.org/10.1128/iai.00238-07" @default.
• W2162007127 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2168294" @default.
• W2162007127 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/17682043" @default.
• W2162007127 hasPublicationYear "2007" @default.
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